• 원예과학기술지
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• 제28권4호
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• pp.648-655
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• 2010

본 연구에서는 들잔디와 돌연변이체에서 Glyphosate 내성 관련 유전자인 EPSPS를 코딩하는 cDNA를 분리하여, 시퀸스 비교분석과 발현 양상 차이 등에 관하여 조사하였다. 5'/3' RACE를 통하여 밝혀진 EPSPS의 cDNA는 각각 1176bp의 open reading frame으로 이루어져 있으며 391개의 아미노산을 코딩하고 있었다. 이는 보고된 다른 EPSPS 유전자들과 높은 유사성을 가지고 있다. Genomic southern 결과 들잔디 내에 EPSPS 유전자는 단일copy로 존재하였다. Wild type과 제초제 내성 변이체의 EPSPS 유전자는 6개의 아미노산 시퀀스의 차이를 보였으며 기존에 보고된 EPSPS active site에서 시퀀스의 차이를 나타냈다. 한편 glyphosate의 독성기작의 하나인 EPSPS 효소활성 저해 작용을 알아본 결과, 내성 개체에서 높은(1.5배 이상) EPSPS 활성을 확인할 수 있었다. Northern 분석과 RT-PCR로 제초제 처리 시간에 따른 EPSPS의 발현량을 살펴본 결과, 제초제 처리 후 시간이 경과할수록 발현량이 증가하다가 7일 이후에는 감소하였고, wild type에 비해 glyphosate 내성 선발개체에서 전체적인 발현량이 높음을 알 수 있었다. 본 연구 결과 선발된 glyphosate 내성 돌연변이체는 높은 EPSPS 효소활성 증가 및 EPSPS active site의 아미노산 시퀀스 변화를 통해 원할하고 지속적인 방향족 아미노산의 합성이 가능한 것으로 보여졌다. 본 실험을 통해 얻어진 glyphosate 내성 잔디 돌연변이체와 방법은 앞으로 유용한 제초제 저항성 돌연변이 품종개발 연구에 활용될 수 있을 것으로 기대된다.

• Journal of Plant Biotechnology
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• 제43권3호
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• pp.311-316
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• 2016

The efficacy of plant breeding has been enhanced by application of molecular markers in population screening and selection. Pepper (Capsicum annuum L.) is a major staple crop that is economically important with worldwide distribution. It is valued for its spicy taste and medicinal effect. The aim of this study was to discover single nucleotide polymorphisms (SNPs), microsatellite markers information, and percentage sharing through orthologous analysis of pepper-specific pungency-related genes. Here, we report the results of transcriptome analysis and microsatellite markers for four pepper varieties that possess a pungency-related gene. Orthologous analyses was performed to identify species-specific pungency-related genes in pepper, Arabidopsis thaliana L., potato (Solanum tuberosum L.), and tomato (Solanum lycopersicum L.). Advancements in next-generation sequencing technologies enabled us to quickly and cost-effectively assemble and characterize genes to select molecular markers in various organisms, including pepper. We identified a total of 9762, 7302, 8596, and 6886 SNPs for the four pepper cultivars Blackcluster, Mandarine, Saengryeg 211, and Saengryeg 213, respectively. We used 454 GS-FLX pyrosequencing to identify microsatellite markers and tri-nucleotide repeats (54.4%), the most common repeats, followed by di-, hexa-, tetra-, and penta-nucleotide repeats. A total of 5156 (15.9%) pepper-specific pungency-related genes were discovered as a result of orthologous analysis.

• Journal of Plant Biotechnology
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• 제35권4호
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• pp.281-290
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• 2008

식물의 Receptor like kinase (RLK) 단백질들은, 그 기능이 잘 알려져 있지 않지만, signal sequence, single transmembrane region, cytoplasmic kinase 도메인으로 구성되어 있다 RLK gene (RLGs)는 호르몬 반응 경로, 세포 분열, 식물의 성장과 발달, 자가 불수분 그리고 공생과 병원균 인식과 연관되어 있다고 보고되었다. 본 연구에서는 벼의 RLGs 중 RLG1, RLC5, RLG6, RLG#6, RLG8, RLG10, RLG17, RLG18, RLG20의 스트레스에 따른 발현 변화를 real-time PCR을 사용하여 분석하여 분자적 특징을 확인하였다. Threshold cycle ($C_T$) 수치의 변화를 통해 발현 변화를 비교하여 결과적으로, NaCl에 대해서는 모두 감소하는 경향을 확인하였으며, Salicylic acid와 상처에 대해서는 각 RLGs에 따라 발현 변화가 다르게 나타났다. 이에 비교하여 저온에서는 모든 RLGs발현이 증가하였다. 따라서 본 연구에서는 냉해에 의한 벼의 생산성 감소를 벼의 RLK에서 확인된 분자적 특징을 이용함으로써 신 기능성 품종의 개발을 통해 극복 할 수 있을 것이라는 가능성을 보여주었다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7097-7103
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• 2014

Over-expression of de novo lipogenesis (DNL) genes is associated with the prognosis of various types of cancers. However, the effects of single nucleotide polymorphisms (SNPs) in these genes on recurrence and survival of non-small cell lung cancer (NSCLC) patients after surgery are still unknown. In this study, a total of 500 NSCLC patients who underwent surgery treatment were included. Eight SNPs in 3 genes (ACACA, FASN and ACLY) of the DNL pathway were examined using the Sequenom iPLEX genotyping system. Multivariate Cox proportional hazards regression and Kaplan-Meier curves were used to analyze the association of SNPs with patient survival and tumour recurrence. We found that two SNPs in the FASN gene were significantly associated with the recurrence of NSCLC. SNP rs4246444 had a significant association with lung cancer recurrence under additive model (hazard ratio [HR], 0.82; 95% confidence interval [95%CI], 0.67-1.00; p=0.05). Under the dominant model, rs4485435 exhibited a significant association with recurrence (HR, 0.75; 95%CI, 0.56-1.01; p=0.05). Additionally, SNP rs9912300 in ACLY gene was significantly associated with overall survival in lung cancer patients (HR, 1.41; 95%CI, 1.02-1.94, p=0.04) under the dominant model. Further cumulative effect analysis showed moderate dose-dependent effects of unfavorable SNPs on both survival and recurrence. Our data suggest that the SNPs in DNL genes may serve as independent prognostic markers for NSCLC patients after surgery.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1256-1264
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• 2016

Adrenergic, alpha-1B-, receptor (ADRA1B) and peroxisome proliferator-activated receptor gamma, coactivator 1 beta (PPARGC1B) genes are involved in regulation of hen ovarian development. In this study, these two genes were investigated as possible molecular markers associated with hen-housed egg production, egg weight (EW) and body weight in Chinese Dagu hens. Samples were analyzed using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique, followed by sequencing analysis. Two novel single nucleotide polymorphisms (SNPs) were identified within the candidate genes. Among them, an A/G transition at base position 1915 in exon 2 of ADRA1B gene and a T/C mutation at base position 6146 in the 3'- untranslated region (UTR) of PPARGC1B gene were found to be polymorphic and named SNP A1915G and T6146C, respectively. The SNP A1915G (ADRA1B) leads to a non-synonymous substitution (aspartic acid 489-to-glycine). The 360 birds from the Dagu population were divided into genotypes AA and AG, allele A was found to be present at a higher frequency. Furthermore, the AG genotype correlated with significantly higher hen-housed egg production (HHEP) at 30, 43, 57, and 66 wks of age and with a higher EW at 30 and 43 wks (p<0.05). For the SNP T6146C (PPARGC1B), the hens were typed into TT and TC genotypes, with the T allele shown to be dominant. The TC genotype was also markedly correlated with higher HHEP at 57 and 66 wks of age and EW at 30 and 43 wks (p<0.05). Moreover, four haplotypes were reconstructed based on these two SNPs, with the AGTC haplotype found to be associated with the highest HHEP at 30 to 66 wks of age and with higher EW at 30 and 43 wks (p<0.05). Collectively, the two SNPs identified in this study might be used as potential genetic molecular markers favorable in the improvement of egg productivity in chicken breeding.

• BMB Reports
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• 제31권6호
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• pp.578-584
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• 1998

In a previous paper (Kim et al., 1996a), the immediate 5' -flanking region and coding region of the human UDP-N -acetylglucosamine:-D-mannoside-1,4-Nacetylglucosaminyltransferase III (N-acetylglucosaminyitransferase- III; GnT-III) gene was reported, isolated and analyzed. Herein, we report on amplification of a new 5' -noncoding region of the GnT-III mRNA by single-strand ligation to single-stranded cDNA-PCR (5' -RACE PCR) using poly(A)+ RNA isolated from human fetal liver cells. A cDNA clone was obtained with 5' sequences (96 bp) that diverged seven nucleotides upstream from the ATG (+1) start codon. A concensus splice junction sequence, TCTCCCGCAG, was found immediately 5' to the position where the sequences of the cDNA diverged. The result suggested the presence of an intron in the 5' -noncoding region and that the cDNA was an incompletely reversetranscribed cDNA product derived from an mRNA containing a new noncoding exon. When mRNA expression of GnT-III in various human tissues and cancer cell lines was examined, Northern blot analysis indicated high expression levels of GnT-III in human fetal kidney and brain tissues, as well as for a number of leukemia and lymphoma cancer cell lines. Promoter activities of the 5' -flanking regions of exon 1 and the new noncoding region were measured in a human hepatoma cell line, HepG2, by luciferase assays. The 5'-flanking region of exon 1 was the most active, whilst that of exon 2 was inactive.

• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1725-1731
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• 2020

Objective: An initial RNA-Sequencing study revealed that UDP-galactose-4-epimerase (GALE) was one of the most promising candidates for milk protein concentration in Chinese Holstein cattle. This enzyme catalyzes the interconversion of UDP-galactose and UDP-glucose, an important step in galactose catabolism. To further validate the genetic effect of GALE on milk protein traits, genetic variations were identified, and genotypes-phenotypes associations were performed. Methods: The entire coding region and the 5'-regulatory region (5'-UTR) of GALE were re-sequenced using pooled DNA of 17 unrelated sires. Association studies for five milk production traits were performed using a mixed linear animal model with a population encompassing 1,027 Chinese Holstein cows. Results: A total of three variants in GALE were identified, including two novel variants (g.2114 A>G and g.2037 G>A) in the 5'-UTR and one previously reported variant (g.3836 G>C) in an intron. All three single nucleotide polymorphisms (SNPs) were associated with milk yield (p<0.0001), fat yield (p = 0.0006 to <0.0001), protein yield (p = 0.0232 to <0.0001) and protein percentage (p<0.0001), while no significant associations were detected between the SNPs and fat percentage. A strong linkage disequilibrium (D' = 0.96 to 1.00) was observed among all three SNPs, and a 5 Kb haplotype block involving three main haplotypes with GAG, AGC, and AGG was formed. The results of haplotype association analyses were consistent with the results of single locus association analysis (p<0.0001). The phenotypic variance ratio above 3.00% was observed for milk protein yield that was explained by SNP-g.3836G >C. Conclusion: Overall, our findings provided new insights into the polymorphic variations in bovine GALE gene and their associations with milk protein concentration. The data indicate their potential uses for marker-assisted breeding or genetic selection schemes.

• 한국자원식물학회지
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• 제34권5호
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• pp.478-490
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• 2021

멜론은 세계 각지에서 재배되는 경제적으로 중요한 작물중의 하나이다. 본 연구는 농업유전자원센터에서 수집 보관중인 멜론 유전자원을 대상으로 다양한 생육 특성을 특성을 조사하고, 멜론의 중요한 육종 형질중의 하나인 과육색의 유전형과 표현형을 조사하여 멜론 육종에 필요한 육종 재료 확보를 위한 기초 자료를 마련하고자 수행되었다. 총 219개의 멜론 유전자원을 대상으로 19개의 생육 특성과 PCA분석을 수행하고, 멜론의 중요한 육종 형질중의 하나인 과육색의 유전형을 조사하여 표현형과 비교하였다. 과육색은 오렌지색, 백색, 녹색, 유백색, 황색의 5가지로 분류하였으며, 이중 오렌지색이 87개로 가장 많았으며, 그 다음으로 백색이 75개였다. 그리고, 오렌지색과 녹색 과육 구별용 마커를 적용한 결과, 녹색 과육 21개의 경우는 표현형과 유전형 일치율이 100%였으며, 오렌지색의 경우는 98%, 백색은 97%, 유백색의 경우는 80%의 일치율을 보였다. 표현형과 유전형이 일치하는 않는 총 8개 유전자원의 염기서열을 분석한 결과, 3곳의 위치에서 단일염기다형성(SNP; single nucleotide polymorphism)이 있었다. 이러한 결과는 멜론의 과육색을 결정하는 아직 알려지지 않은 유전기작이 존재한다는 것을 제시하였으며, 본 연구에서 얻어진 다양한 유전자원의 생육조사 결과는 멜론 육종에 유용하게 쓰일 것으로 생각된다.

• 대한임상검사과학회지
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• 제53권3호
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• pp.249-256
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• 2021

천식은 만성 염증성 기도 폐쇄 질환이다. 질병 발생 요인은 다양하며 특히, 유전적 요인과 환경적 요인이 천식 발병에 영향을 미치는 것으로 추정된다. MAPK (mitogen-activated protein kinase)경로는 Th1/Th2의 균형을 조절하며, 천식 발생에 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 MAPK 경로를 조절하는 NDFIP2 유전자와 천식 발병과의 상관관계를 분석하였다. 193건의 천식 환자와 3,228건의 정상 대조군의 유전형 데이터를 사용하였다. 그 결과 NDFIP2 안에 있는 4개의 SNP이 천식과 유의한 상관관계와 높은 상대적 위험도를 보였다. 특히 NDFIP2의 rs2783122는 천식과 통계적으로 가장 유의한 연관성을 나타냈다(P-value=9.76×10^-6, OR=1.67, 95% CI=1.33~2.10). NDFIP2 유전자에 대한 SNP imputation 결과 16개의 SNP가 추가 발견되었으며, 모두 유의한 상관 관계와 높은 상대적 위험도를 나타냈다. 유전자형 기반 mRNA 발현 분석을 통해 rs1408049가 minor allele을 가질 경우 유전자 발현이 증가됨을 알 수 있었다. 증가된 NDFIP2 발현은 MAPK 경로를 활성화시켜 천식 발병에 영향을 미칠 수 있다. 결론적으로 NDFIP2의 다형성은 천식 발병과 관련이 있으며, 이는 한국 인구의 천식 관리에 대한 새로운 지침을 제공할 수 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3629-3633
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• 2014

Background: Glioblastoma (GBM) is an immunosuppressive tumor whose median survival time is only 12-15 months, and patients with GBM have a uniformly poor prognosis. It is known that heredity contributes to formation of glioma, but there are few genetic studies concerning GBM. Materials and Methods: We genotyped six tagging SNPs (tSNP) in Han Chinese GBM and control patients. We used Microsoft Excel and SPSS 16.0 statistical package for statistical analysis and SNP Stats to test for associations between certain tSNPs and risk of GBM in five different models. ORs and 95%CIs were calculated for unconditional logistic-regression analysis with adjustment for age and gender. The SHEsis software platform was applied for analysis of linkage disequilibrium, haplotype construction, and genetic associations at polymorphism loci. Results: We found rs891835 in CCDC26 to be associated with GBM susceptibility at a level of p=0.009. The following genotypes of rs891835 were found to be associated with GBM risk in four different models of gene action: i) genotype GT (OR=2.26; 95%CI, 1.29-3.97; p=0.019) or GG (OR=1.33; 95%CI, 0.23-7.81; p=0.019) in the codominant model; ii) genotypes GT and GG (OR=2.18; 95%CI, 1.26-3.78; p=0.0061) in the dominant model; iii) GT (OR=2.24; 95%CI, 1.28-3.92; p=0.0053) in the overdominant model; iv) the allele G of rs891835 (OR=1.85; 95%CI, 1.14-3.00; p=0.015) in the additive model. In addition, "CG" and "CGGAG" were found by haplotype analysis to be associated with increased GBM risk. In contrast, genotype GG of CCDC26 rs6470745 was associated with decreased GBM risk (OR=0.34; 95%CI, 0.12-1.01; p=0.029) in the recessive model. Conclusions: Our results, combined with those from previous studies, suggest a potential genetic contribution of CCDC26 to GBM progression among Han Chinese.